Genetically modified seed combined with spore forming bacterium and optional insect control agents and methods for treating plants

ABSTRACT

Products are provided that improve overall plant vigor and yield by combining agriculturally effective amounts of at least one spore-forming bacterium and at least one optional insect control agent to a genetically modified plant, plant part, or seed. This product is particularly effective in the presence of plant parasitic nematode and fungal species. Use of the product leads to an overall reduction in crop losses caused by either plant parasitic nematodes or fungi and this reduction is much greater than using genetically modified seed with just an insect control agent. According to some embodiments, the use of the product results in about a 2%-10% increase in soybean bushel yield, 3%-6.5% increase in cotton yield, and 3%-8% in corn bushel yield. Methods for utilizing and manufacturing the combination are also provided.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/848,873, filed Sep. 9, 2015, which is a continuation application ofU.S. application Ser. No. 13/059,124, filed Mar. 2, 2011, which is a §371 National Stage Application of PCT/US09/55842, filed Sep. 3, 2009,which claims priority to U.S. Provisional Application No. 61/191,620,filed Sep. 10, 2008, the contents of which are incorporated herein byreference in their entireties.

BACKGROUND OF THE INVENTION 1. Field of the Invention

The invention relates generally to products and methods for reducingoverall damage and losses in plant health, vigor, and yield caused byplant parasitic nematode and fungi. More specifically, the inventionrelates to products comprising genetically modified seed and at leastone agriculturally beneficial spore-forming bacterium combined with anoptional insect control agent, and methods for utilizing the combinationfor treating genetically modified seeds, plants and plant parts.

2. Description of Related Art

Nematodes are microscopic unsegmented worms known to reside in virtuallyevery type of environment (terrestrial, freshwater, marine). Of the over80,000 known species, many are agriculturally significant, particularlythose classified as pests. One such species is the root knot nematodewhich attacks a broad range of plants, shrubs, and crops. Thesesoil-born nematodes attack newly formed roots causing stunted growth,swelling or gall formation. The roots may then crack open thus exposingthe roots to other microorganisms such as bacteria and fungi. Withenvironmentally friendly practices such as reduced or no tillagefarming, and various nematode species acquiring resistance to transgenicseed, nematode related crop losses appear to be on the rise.

Chemical nematicides such as soil fumigants or non-fumigants have beenin use for many years to combat infestations. Such nematicides mayrequire repeated applications of synthetic chemicals to the ground priorto planting. Due to their toxicity, chemical nematicides have come underscrutiny from the Environmental Protection Agency (EPA) and in somecases their use has been limited or restricted by the EPA As the use oftraditional chemical nematicides such as methyl-bromide andorganophosphates continue to be phased out, a need for the developmentof alternative treatment options has arisen. U.S. Pat. No. 6,593,273discloses treatment of transgenic corn seeds with pesticides to treatnematode infestations.

U.S. Pat. No. 6,844,339 discloses using a neonicotinoid to controlnematodes. The preferred compounds in the '339 patent are nitroimino ornitroguanidino compounds. The neonicotinoid can be applied to either thenematode environment or plant material itself. WO/2007/149817 disclosescombining a biological control agent with a nematicide, such asavermectin, to enhance plant protection against pests and pathogens.This combination, however, does not address the toxicity of usingcertain chemical nematicides.

SUMMARY OF THE INVENTION

There remains a need for effective compositions and methods that useenvironmentally friendly biological components and less toxic chemicalnematicides, but utilize them in such a manner that they can provideimproved plant vigor and yield without the use of more toxic traditionalchemical nematicides.

The invention provides improved products and methods for controllingnematode damage or infestations. The product uses at least one sporeforming bacterium and an optional insect control agent an optionalfungicide control agent in combination with a genetically modified seed,plant, or plant part.

Methods for treating a seed, plant and/or plant part are also provided.The method comprises (a) providing a composition comprising an effectiveamount of at least one spore-forming bacterium; (b) combining thespore-forming bacterium with an optional insect control agent; and (c)applying the composition to the genetically modified seed, plant, and/orplant part. Application can be done in any desired manner, such as inthe form of seed coating, soil drench, and/or directly in-furrow and/oras a foliar spray and applied either pre-emergence, post-emergence orboth. Optionally, the insect control agent can be applied separately tothe genetically modified seed, plant, or plant part. Further, at leastone fungicide may also be combined with the spore-forming bacterium,optional insect control agent, or applied separately to the geneticallymodified plant, seed, or plant part. In sum, the individual componentsor composition can be applied to the seed, the plant, the plant foliar,to the fruit of the plant, or the soil wherein the plant is growing orwherein it is desired to grow.

According to one aspect of the invention, a product is providedcomprising a spore-forming bacterium combined with an optional insectcontrol agent and a genetically modified seed.

In another aspect of the invention, a method of treating a geneticallymodified seed, plant, or plant part is provided, comprising applying tothe seed, plant or plant part at least one spore-forming bacterium; and,optionally, at least one insect control agent.

In a further aspect of the invention, a method of protecting agenetically modified seed, plant, or plant part from nematodes isprovided, comprising providing at least one composition comprising0.0001 to 20% by weight of at least one spore-forming bacterium and0.001 to 20% by weight of at least one insect control agent; andapplying the composition to the seed, plant, or plant part.

In yet another aspect of the invention, a composition for protecting agenetically modified seed, plant, or plant part from nematodes isprovided, comprising: (i) at least one spore-forming bacterium in anamount of from about 2% by weight to 80% by weight; (ii) at least oneinsect control agent in an amount of from about 1% by weight to about80% by weight; and (iii) a solvent.

In yet a further aspect of the invention, a method of manufacturing agenetically modified seed treated with at least one spore-formingbacteria and an optional insect control agent is provided, comprising:(i) applying said spore-forming bacteria and optional insect controlagent to said genetically modified seed; and (ii) mixing saidgenetically modified seed to achieve a substantially uniform treatment.

Other products and methods in accordance with the composition areprovided in the detailed description and claims that follow below.Additional objects, features, and advantages will be sent forth in thedescription that follows, and in part, will be obvious from thedescription, or may be learned by practice of the invention. Theobjects, features, and advantages may be realized and obtained by meansof the instrumentalities and combination particularly pointed out in theappended claims.

DESCRIPTION Detailed Description of the Invention

The products disclosed herein have been found to provide a greaterdegree of plant vigor and yield in nematode infested environments thanwould be expected from the application of insecticides to geneticallymodified seeds, plants, and plant parts. At least some of the optionalinsect control agents have been shown to provide increased root masseven in the absence of insect pressure which increased root mass leadsto improved establishment of the beneficial bacteria within therhizosphere which, in turn, reduces overall losses in crop vigor andyields caused by either plant parasitic nematodes or fungi. Along withthe physical combination of these components while treating plants andplant parts, the compositions may be formulated to provide a stableenvironment for living spore-forming bacteriums such as spore-forming,root-colonizing bacteria. Various additives, such as fungicides,insecticides, stabilizers, emulsifiers, may be added to thespore-forming bacterium and/or genetically modified seed, plant, orplant part depending on the desired properties.

The genetically modified seeds, plants, or plant parts are typicallydeveloped for insect control and herbicide tolerance. Thus, the additionof utilizing a spore forming bacterium with the optional insect controlagent helps complete the ability of the seed to survive under adverseconditions. The at least one spore-forming bacterium generally hasproven agriculturally beneficial to colonize a plant's root system. Theoptional insect control agent can be at least one chemical insecticidethat, whether or not having proven direct nematicidal or fungicidalactivity, does possess the proven ability to increase the mass of theplant's root system to which it is applied. The genetically modifiedseed can be any seed that results in a genetically modified plant orplant part that expresses insect toxins or herbicide resistance.Further, the genetically modified seed can be any seed that results in agenetically modified plant or plant part that expresses toxins orresistance to bacterial and fungi. Moreover, the genetically modifiedseed may be any seed that results in a genetically modified plant orplant part that expresses tolerance to environmental factors such aswater stress and nitrogen production.

Regarding insect toxins, U.S. Pat. No. 5,877,012, herein incorporated byreference in its entirety, discloses the cloning and expression ofproteins from such organisms as Bacillus, Pseudomonas, Clavibacter, andRhizobium into plants to obtain transgenic plants with resistance tosuch pests as black cutworms, armyworms, and borers. Further, U.S. Pat.Nos. 5,625,136 and 5,859,336, hereby incorporated by reference in theirentirety, disclose transforming corn plants with a gene from B.thuringiensis that encodes for delta-endotoxins proving the transgeniccorn with improved resistance to European corn borers. A comprehensivereport of field trials of transgenic corn that expresses an insecticidalprotein from B. thuringiensis has been provided by Armstrong et al., inCrop Science, 35(2):550-557 (1995), hereby incorporated by reference inits entirety. Additional references that disclose corn encoded with theB. thuringiensis gene, include U.S. Pat. Nos. 4,766,203; 4,797,279; and4,910,016, hereby incorporated by reference in their entirety; and WO99/312248, hereby incorporated by reference in its entirety. Regardingherbicide resistance, U.S. Pat. No. 4,971,908, herein incorporated byreference in its entirety, discloses genetically modified plants thatare glyphosate resistant. Glyphosate resistance is achieved bygenetically modifying the plant or seed to produce mutant EPSP synthaseenzymes that exhibit a lower affinity for glyphosate while maintainingcatalytic activity. Additional references that disclose glyphosateresistant plants and seeds include U.S. Pat. Nos. 5,463,175; 5,776,760;5,804,425; 6,689,880; 6,803,501; 7,214,535; and 7,335,816, hereinincorporated by reference in their entirety.

The one spore-forming bacterium has demonstrated agriculture benefit.Preferably, the at least one spore-forming bacteria is a root colonizingbacterium (e.g. rhizobacterium). Agriculture benefit refers to thebacterium's ability to provide protection from the harmful effects ofplant pathogenic fungi or bacteria and/or soil born animals such asthose belonging to the phylum Nematoda or Aschelminthes. Protectionagainst plant parasitic nematodes and parasitic microorganisms can occurthrough chitinolytic, proteolytic, collagenolytic, or other activitiesdetrimental to these soil born animals and/or detrimental to microbialpopulations. Bacteria exhibiting these nematicidal, fungicidal andbactericidal properties may include but are not limited to, Bacillusargri, Bacillus aizawai, Bacillus albolactis, Bacillusamyloliquefaciens, Bacillus cereus, Bacillus coagulans, Bacillusendoparasiticus, Bacillus endorhythmos, Bacillus firmus, Bacilluskurstaki, Bacillus lacticola, Bacillus lactimorbus, Bacillus lactis,Bacillus laterosporus, Bacillus lentimorbus, Bacillus licheniformis,Bacillus megaterium, Bacillus medusa, Bacillus metiens, Bacillus natto,Bacillus nigrificans, Bacillus popillae, Bacillus pumilus, Bacillussiamensis, Bacillus sphearicus, Bacillus spp., Bacillus subtilis,Bacillus thurngiensis, Bacillus unifagellatus, plus those listed in thecategory of Bacillus Genus in Bergey's Manual of SystematicBacteriology, First Ed. (1986), hereby incorporated by reference in itsentirety.

Preferably, the spore-forming bacterium is at least one B. firmus CNCM1-1582 spore and/or B. cereus strain CNCM I-1562 spore as disclosed inU.S. Pat. No. 6,406,690, hereby incorporated by reference in itsentirety. Most preferably, the spore-forming bacterium is B. firmus CNCMI-1582. Alternatively, the spore-forming bacterium can be at least oneB. amyloliquefaciens IN937a, at least one Bacillus subtillis straindesignated GB03, or at least one B. pumulis strain designated GB34.Further, the spore-forming bacterium can be a mixture of any specieslisted above, as well as other spore-forming, root colonizing bacteriaknown to exhibit agriculturally beneficial properties.

In a preferred embodiment, the spore-forming bacterium can be applied tothe seed, plant, or plant parts as either a powder, aqueous, ornon-aqueous solution. Powders can be either dry, wettable powders, orwater dispersable granules. Preferably, the spore-forming bacterium is asolution, emulsifiable concentrate, wettable powder, suspensionconcentrate, soluble powder, granules, suspension-emulsion concentrate,natural and synthetic materials impregnated with active compounds, andfine control release capsules. The spore-forming bacterium in a liquidor dry form may be admixed with the soil prior to, at the time of, orafter planting. Most preferably, the formulation is in a liquid stateadmixed with the soil prior to or at the time of planting.

The amount of the at least one spore-forming bacterium employed in thecompositions can vary depending on the final formulation as well as sizeor type of the plant, plant part, or seed to be utilized. Preferably, atleast one spore-forming bacterium in the compositions is present inabout 2% by weight of total formulation to about 80% by weight of totalformulation. More preferably, about 5% by weight of total formulation toabout 65% by weight of total formulation; and most preferably about 10%by weight of total formulation to about 60% by weight of totalformulation.

The compositions further comprise at least one optional insect controlagent. Preferably, the insect control agent can be any insecticidalchemical compound or composition having insecticidal activity, but nodirect nematicidal activity and no detrimental activity against theutilized spore-forming bacterium, and preferably also has the addedability to increase root mass upon application. Alternatively, thecombination may comprise at least one additional chemical compound thatdoes exhibit nematicidal or fungicidal properties. Such compositions canby useful in geographical areas having extremely high populations ofnematode infestation or to provide additional fungicidal activityagainst heavy fungal disease pressure. The plant, seed, or plantmaterial can be treated separately or simultaneously with the additionalinsect or fungicidal control agent. Most preferably, the insect controlagent is a non-nematicidal neonicotinoid insecticide compound of formula(I)

In another preferred embodiment, the optional insect control agent is atleast one systemic, non-nematicidal neonicotinoid insecticide compoundof formula (I)

wherein

-   -   A is 2-chloropyrid-5-yl, 2-methylpyrid-5-yl,        1-oxido-3-pyridinio, 2-chloro-1-oxido-5-pyridinio,        2,3-dichloro-1-oxido-5-pyridinio, tetrahydrofuran-3-yl,        5-methyl-tetrahydrofuran-3-yl or 2-chlorothiazol-5-yl group;    -   R is hydrogen, C.sub.1-C.sub.6 alkyl, phenyl-C.sub.1-C.sub.4        alkyl, C.sub.3-C.sub.6 cycloalkyl, C.sub.2-C.sub.6 alkenyl or        C.sub.2-C.sub.6 alkynyl;    -   Y is —N(R)(R.sub.2) or SR.sub.2;    -   R.sub.1 and R.sub.2 are independently of each other        C.sub.1-C.sub.4-alkyl, C.sub.1-C.sub.4-alkenyl,        C.sub.1-C.sub.4-alkinyl, —C(═O)—CH.sub.3 or benzyl; or together        form a group —CH.sub.2-CH.sub.2-, —CH.sub.2-CH.sub.2CH.sub.2-,        CH.sub.2-O—CH.sub.2CH.sub.2-S—CH.sub.2-, CH.sub.2-NH—CH.sub.2-        or —CH.sub.2-N(CH.sub.3)-CH.sub.2-; and    -   X is N—NO.sub.2 or N—CN or CH—NO.sub.2.

Particularly preferred non-nematicidal, neonicotinoid insecticidesinclude 1-(6-chloro-3-pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine(imidacloprid),3-(6-chloro-3-pyridylmethyl)-1,3-thiazolidin-2-ylidenecyanamide(thiacloprid),1-(2-chloro-1,3-thiazol-5-ylmethyl)-3-methyl-2-nitroguanidine(clothianidin), nitempyran,N.sup.1-[(6-chloro-3-pyridyl)methyl]-N.sup.2-cyano-N.sup.1-methylacetamidine(acetamiprid),3-(2-chloro-1,3-thiazol-5-ylmethyl)-5-methyl-1,3,5-oxadiazinan-4-ylidene(nitro)amine(thiamethoxam) and1-methyl-2-nitro-3-(tetrahydro-3-furylmethyl)guanidine (dinotefuran).

In an alternative embodiment, the spore-forming bacterium—insect controlagent combination can optionally include an additional chemical compoundwith direct nematicidal activity. Suitable nematicidal insect controlagents include antibiotic nematicides such as abamectin; carbamatenematicides such as benomyl, carbofuran, carbosulfan, and cleothocard;oxime carbamate nematicides such as alanycarb, aldicarb, aldoxycarb,oxamyl; organophosphorous nematicides such as diamidafos, fenamiphos,fosthietan, phosphamidon, cadusafos, chlorpyrifos, diclofenthion,dimethoate, ethoprophos, fensulfothion, fostiazate, heterophos,isamidofos, isazofos, methomyl, phorate, phosphocarb, terbufos,thiodicarb, thionazin, triazophos, imicyafos, and mecarphon. Othersuitable nematicidal insect control agents include acetoprole,benclothiaz, chloropicrin, dazomet, DBCP, DCIP, 1,2-dichloropropane,1,3-dichloropropene, furfural, iodomethane, metam, methyl bromide,methyl isothiocyanate, and xylenols. Alternatively, the spore-formingbacterium can also be combined with biological nematicide agents such asMyrothecium verrucaria, Burholderia cepacia, Bacillus chitonosporus andPaecilomyces lilacinus or nematicidal agents of plant or animal originsuch as harpin proteins, amino acid sequences or virus, viroidparticles.

The amount of the at least one optional insect control agent added tothe spore-forming bacterium can vary depending on the final formulationas well as the size of the plant and seed to be treated. Preferably, theat least one insect control agent is about 1% by weight of totalformulation to about 80% by weight of total formulation. Morepreferably, the insect control agent is present in an amount of about 5%by weight of total formulation to about 60% by weight of totalformulation. Most preferably, the insect control agent is present in anamount of about 10% by weight of total formulation to about 50% byweight of total formulation.

In a further embodiment, the spore-forming bacterium, alone or incombination with the insect control agent, can further comprise aneffective amount of at least one fungicide. Preferred fungicides includealdimorph, ampropylfos, ampropylfos potassium, andoprim, anilazine,azaconazole, azoxystrobin, benalaxyl, benodanil, benomyl, benzamacril,benzamacryl-isobutyl, bialaphos, binapacryl, biphenyl, bitertanol,blasticidin-S, boscalid, bromuconazole, bupirimate, buthiobate, calciumpolysulphide, capsimycin, captafol, captan, carbendazim, carboxin,carvon, quinomethionate, chlobenthiazone, chiorfenazole, chloroneb,chloropicrin, chlorothalonil, chlozolinate, clozylacon, cufraneb,cymoxanil, cyproconazole, cyprodinil, cyprofuram, debacarb,dichlorophen, diclobutrazole, diclofluanid, diclomezine, dicloran,diethofencarb, difenoconazole, dimethirimol, dimethomorph,dimoxystrobin, diniconazole, diniconazole-M, dinocap, diphenylamine,dipyrithione, ditalimfos, dithianon, dodemorph, dodine, drazoxolon,edifenphos, epoxiconazole, etaconazole, ethirimol, etridiazole,famoxadon, fenapanil, fenarimol, fenbuconazole, fenfuram, fenitropan,fenpiclonil, fenpropidin, fenpropimorph, fentin acetate, fentinhydroxide, ferbam, ferimzone, fluazinam, fludioxonil, flumetover,fluoromide, fluquinconazole, flurprimidol, flusilazole, flusulfamide,flutolanil, flutriafol, folpet, fosetyl-aluminium, fosetyl-sodium,fthalide, fuberidazole, furalaxyl, furametpyr, furcarbonil, furconazole,furconazole-cis, furmecyclox, guazatine, hexachlorobenzene,hexaconazole, hymexazole, imazalil, imibenconazole, iminoctadine,iminoctadine albesilate, iminoctadine triacetate, iodocarb, ipconazole,iprobenfos (IBP), iprodione, irumamycin, isoprothiolane, isovaledione,kasugamycin, kresoxim-methyl, copper preparations, such as: copperhydroxide, copper naphthenate, copper oxychloride, copper sulphate,copper oxide, oxine-copper and Bordeaux mixture, mancopper, mancozeb,maneb, meferimzone, mepanipyrim, mepronil, metalaxyl, metconazole,methasulfocarb, methfuroxam, metiram, metomeclam, metsulfovax,mildiomycin, myclobutanil, myclozolin, nickel dimethyldithiocarbamate,nitrothal-isopropyl, nuarimol, ofurace, oxadixyl, oxamocarb, oxolinicacid, oxycarboxim, oxyfenthiin, paclobutrazole, pefurazoate,penconazole, pencycuron, phosdiphen, pimaricin, piperalin, polyoxin,polyoxorim, probenazole, prochloraz, procymidone, propamocarb,propanosine-sodium, propiconazole, propineb, prothiocinazole,pyraclostrobin, pyrazophos, pyrifenox, pyrimethanil, pyroquilon,pyroxyfur, quinconazole, quintozene (PCNB), sulphur and sulphurpreparations, tebuconazole, tecloftalam, tecnazene, tetcyclasis,tetraconazole, thiabendazole, thicyofen, thifluzamide,thiophanate-methyl, thiram, tioxymid, tolclofos-methyl, tolylfluanid,triadimefon, triadimenol, triazbutil, triazoxide, trichlamide,tricyclazole, tridemorph, trifloxystrobin, triflumizole, triforine,triticonazole, uniconazole, validamycin A, vinclozolin, viniconazole,zarilamide, zineb, ziram and also Dagger G, OK-8705, OK-8801,α-(1,1-dimethylethyl)-β-(2-phenoxyethyl)-1H-1,2,4-triazole-1-ethanol,α-(2,4-dichlorophenyl)-β-fluoro-β-propyl-1H-1,2,4-triazole-1-ethanol,α-(2,4-dichlorophenyl)-β-methoxy-α-methyl-1H-1,2,4-triazole-1-ethanol,α-(5-methyl-1,3-dioxan-5-yl)-β-[[4-(trifluoromethyl)-phenyl]-methylene]-1H-1,2,4-triazole-1-ethanol,(5RS,6RS)-6-hydroxy-2,2,7,7-tetramethyl-5-(1H-1,2,4-triazol-1-yl)-3-octanone,(E)-α-(methoxyimino)-N-methyl-2-phenoxy-phenylacetamide,1-isopropyl{2-methyl-1-[[[1-(4-methylphenyl)-ethyl]-amino]-carbonyl]-propyl}carbamate,1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-ethanone-O-(phenylmethyl)-oxime,1-(2-methyl-1-naphthalenyl)-1H-pyrrole-2,5-dione,1-(3,5-dichlorophenyl)-3-(2-propenyl)-2,5-pyrrolidindione,1-[(diiodomethyl)-sulphonyl]-4-methyl-benzene,1-[[2-(2,4-dichlorophenyl)-1,3-dioxolan-2-yl]-methyl]-1H-imidazole,1-[[2-(4-chlorophenyl)-3-phenyloxiranyl]-methyl]-1H-1,2,4-triazole,1-[1-[2-[(2,4-dichlorophenyl)-methoxy]-phenyl]-ethenyl]-1H-imidazole,1-methyl-5-nonyl-2-(phenylmethyl)-3-pyrrolidinole,2′,6′-dibromo-2-methyl-4′-trifluoromethoxy-4′-trifluoro-methyl-1,3-thiazole-5-carboxanilide,2,2-dichloro-N-[1-(4-chlorophenyl)-ethyl]-1-ethyl-3-methyl-cyclopropanecarboxamide,2,6-dichloro-5-(methylthio)-4-pyrimidinyl-thiocyanate,2,6-dichloro-N-(4-trifluoromethylbenzyl)-benzamide,2,6-dichloro-N-[[4-(trifluoromethyl)-phenyl]-methyl]-benzamide,2-(2,3,3-triiodo-2-propenyl)-2H-tetrazole,2-[(1-methylethyl)-sulphonyl]-5-(trichloromethyl)-1,3,4-thiadiazole,2-[[6-deoxy-4-O-(4-O-methyl-((β-D-glycopyranosyl)-α-D-glucopyranosyl]-amino]-4-methoxy-1H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile,2-aminobutane, 2-bromo-2-(bromomethyl)-pentanedinitrile,2-chloro-N-(2,3-dihydro-1,1,3-trimethyl-1H-inden-4-yl)-3-pyridinecarboxamide,2-chloro-N-(2,6-dimethylphenyl)-N-(isothiocyanatomethyl)-acetamide,2-phenylphenol (OPP),3,4-dichloro-1-[4-(difluoromethoxy)-phenyl]-1H-pyrrole-2,5-dione,3,5-dichloro-N-[cyano[(1-methyl-2-propynyl)-oxy]-methyl]-benzamide,3-(1,1-dimethylpropyl-1-oxo-1H-indene-2-carbonitrile,3-[2-(4-chlorophenyl)-5-ethoxy-3-isoxazolidinyl]-pyridine,4-chloro-2-cyano-N,N-dimethyl-5-(4-methylphenyl)-1H-imidazole-1-sulphonamide,4-methyl-tetrazolo[1,5-a]quinazolin-5(4H)-one,8-(1,1-dimethylethyl)-N-ethyl-N-propyl-1,4-dioxaspiro[4,5]decane-2-methanamine,8-hydroxyquinoline sulphate,9H-xanthene-2-[(phenylamino)-carbonyl]-9-carboxylic hydrazide,bis-(1-methylethyl)-3-methyl-4-[(3-methylbenzoyl)-oxy]-2,5-thiophenedicarboxylate,cis-1-(4-chlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-cycloheptanol,cis-4-[3-[4-(1,1-dimethylpropyl)-phenyl-2-methylpropyl]-2,6-dimethyl-morpholinehydrochloride, ethyl [(4-chlorophenyl)-azo]-cyanoacetate, potassiumbicarbonate, methanetetrathiol-sodium salt, methyl1-(2,3-dihydro-2,2-dimethyl-1H-inden-1-yl)-1H-imidazole-5-carboxylate,methyl N-(2,6-dimethylphenyl)-N-(5-isoxazolylcarbonyl)-DL-alaninate,methyl N-(chloroacetyl)-N-(2,6-dimethylphenyl)-DL-alaninate,N-(2,3-dichloro-4-hydroxyphenyl)-1-methyl-cyclohexanecarboxamide,N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-furanyl)-acetamide,N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-thienyl)-acetamide,N-(2-chloro-4-nitrophenyl)-4-methyl-3-nitro-benzenesulphonamide,N-(4-cyclohexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidinamine,N-(4-hexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidinamine,N-(5-chloro-2-methylphenyl)-2-methoxy-N-(2-oxo-3-oxazolidinyl)-acetamide,N-(6-methoxy)-3-pyridinyl)-cyclopropanecarboxamide,N-[2,2,2-trichloro-1-[(chloroacetyl)-amino]-ethyl]-benzamide,N-[3-chloro-4,5-bis(2-propinyloxy)-phenyl]-N′-methoxy-methanimidamide,N-formyl-N-hydroxy-DL-alanine-sodium salt,0,0-diethyl[2-(dipropylamino)-2-oxoethyl]-ethylphosphoramidothioate,O-methyl S-phenyl phenylpropylphosphoramidothioate, S-methyl1,2,3-benzothiadiazole-7-carbothioate, andspiro[2H]-1-benzopyrane-2,1′(3′H)-isobenzofuran]-3′-one, alone or incombination.

The methods disclosed herein have been found to provide a greater degreeof plant vigor and yield in nematode infested environments than would beexpected from the application of insecticides alone to geneticallymodified seeds, plants, or plant parts. Various additives, such asfungicides, insecticides, stabilizers, emulsifiers, may be applied tothe genetically modified seed or plant depending on the desiredproperties.

The methods include applying at least one spore-forming bacteriumcombined with an optional insect control agent, and optional fungicidecontrol agent, to a genetically modified seed, plant, or plant part.Preferably, the spore-forming bacterium is in solution form,emulsifiable concentrate, wettable powders, suspension concentrate,soluble powders, granules, suspension-emulsion concentrate, natural andsynthetic materials impregnated with active compound, and fine controlrelease capsules in polymeric substances. Preferably, the insect controlagent, if present, is mixed with the spore forming bacterium, andapplied simultaneously with the spore forming bacterium. Optionally, theinsect control agent can be applied separately to the seed, plant, orplant part. Further, if a fungicide control agent is present, this maybe combined with the spore-forming bacterium/insect control agent, orapplied separately. If the spore-forming bacterium and insect controlagent are in powder form, they may be applied directly to the soil,seed, or foliar separately or mixed together at the time of use. If inliquid form, the spore-forming bacterium and insect control agent, maybe sprayed or atomized foliarly or in-furrow at the time of planting,either separately or mixed together at the time of treating.Alternatively, the liquid combination can be introduced to the soilbefore germination of the seed or directly to the soil in contact withthe roots by utilizing a variety of techniques included, but not limitedto, drip irrigation, sprinklers, soil injection or soil drenching.Preferably, the liquid is applied to the seed before planting.

Depending on the final formulation and method of application, one ormore suitable additives can also be introduced to the spore-formingbacterium and combinations thereof. Additives such ascarboxymethylcellulose and natural and synthetic polymers in the form ofpowders, granules, or latexes, such as gum Arabic, chitin, polyvinylalcohol and polyvinyl acetate, as well as natural phospholipids, such ascephalins and lecithins, and synthetic phospholipids, can be added tothe present compositions.

In a preferred embodiment, the spore-forming bacterium, optional insectcontrol agent, and optional fungicide control agent, are formulated in asingle, stable solution, or emulsion or suspension. For solutions, theactive chemical compounds (insect control agents and optional fungicidecontrol agent) are dissolved in solvents before adding the spore-formingbacterium. Suitable liquid solvents include petroleum based aromatics,such as xylene, toluene, or alkylnapthalenes, aliphatic hydrocarbons,such as cyclohexane or paraffins, for example petroleum fractions,mineral and vegetable oils, alcohols, such as butanol or glycol as wellas their ethers and esters, ketones, such as methyl ethyl ketone, methylisobutyl ketone or cyclohexanone, strongly polar solvents, such asdimethylformamide and dimethyl sulphoxide. For emulsions andsuspensions, the liquid medium or solvent is water. The spore-formingbacterium, optional insect control agent, and optional fungicide controlagent may be suspended in separate liquids and mixed at the time ofapplication. Preferably, the spore forming bacterium, optional insectcontrol agent, and optional fungicide control agent, are combined in aready to use formulation that exhibits a shelf-life of preferably twoyears. In use, the liquid can be sprayed or atomized foliarly orin-furrow at the time of planting the corp. The liquid composition canbe introduced to the soil before germination of the seed or directly tothe soil in contact with the roots by utilizing a variety of techniquesincluding, but not limited to, drip irrigation, sprinklers, soilinjection or soil drenching.

Optionally, stabilizers and buffers can be added, including alkaline andalkaline earth metal salts and organic acids, such as citric acid andascorbic acid, inorganic acids, such as hydrochloric acid or sulfuringacid. Biocides can also be added and can included formaldehydes orformaldehyde-releasing agents and derivatives of benzoic acid, such asp-hydroxybenzoic acid. Further additives include functional agentscapable of protecting seeds from harmful effects of selective herbicidessuch as activated carbon, nutrients (fertilizers), and other agentscapable of improving the germination and quality of the products or acombination thereof.

Preferably, the spore-forming bacterium and optional insect controlagents are formulated as a liquid seed treatment. The seed treatmentcomprises at least one spore forming bacterium, and at least oneoptional insect control agent. Optionally, a fungicide control agent canbe mixed with the spore-forming bacterium and insect control agent. Theseeds are substantially uniformly coated with one or more layers ofspore-forming bacterium, optional insect control agent, and optionalfungicide control agent, using conventional methods of mixing, sprayingor a combination thereof. Application is done using specificallydesigned and manufactured equipment that accurately, safely, andefficiently applies seed treatment products to seeds. Such equipmentuses various types of coating technology such as rotary coaters, drumcoaters, fluidized bed techniques, spouted beds, rotary mists or acombination thereof. Preferably, the application is done via either aspinning “atomizer” disk or a spray nozzle which evenly distributes theseed treatment onto the seed as it moves through the spray pattern.Preferably, the seed is then mixed or tumbled for an additional periodof time to achieve additional treatment distribution and drying. Theseeds can be primed or unprimed before coating with the inventivecompositions to increase the uniformity of germination and emergence. Inan alternative embodiment, a dry powder composition can be metered ontothe moving seed.

The seeds may be coated via a continuous or batch coating process. In acontinuous coating process, continuous flow equipment simultaneouslymeters both the seed flow and the seed treatment products. A slide gate,cone and orifice, seed wheel, or weight device (belt or diverter)regulates seed flow. Once the seed flow rate through treating equipmentis determined, the flow rate of the seed treatment is calibrated to theseed flow rate in order to deliver the desired dose to the seed as itflows through the seed treating equipment. Additionally, a computersystem may monitor the seed input to the coating machine, therebymaintaining a constant flow of the appropriate amount of seed.

In a batch coating process, batch treating equipment weighs out aprescribed amount of seed and places the seed into a closed treatingchamber or bowl where the corresponding of seed treatment is thenapplied. The seed and seed treatment are then mixed to achieve asubstantially uniform coating on each seed. This batch is then dumpedout of the treating chamber in preparation for the treatment of the nextbatch. With computer control systems, this batch process is automatedenabling it to continuously repeat the batch treating process.

In either coating process, the seed coating machinery can optionally beoperated by a programmable logic controller that allows variousequipment to be started and stopped without employee intervention. Thecomponents of this system are commercially available through severalsources such as Gustafson Equipment of Shakopee, Minn.

A variety of additives can be added to the seed treatments. Binders canbe added and include those composed preferably of an adhesive polymerthat can be natural or synthetic without phytotoxic effect on the seedto be coated. A variety of colorants may be employed, including organicchromophores classified as nitroso, nitro, azo, including monoazo,bisazo, and polyazo, diphenylmethane, triarylmethane, xanthene, methane,acridine, thiazole, thiazine, indamine, indophenol, azine, oxazine,anthraquinone, and phthalocyanine. Other additives that can be addedinclude trace nutrients such as salts of iron, manganese, boron, copper,cobalt, molybdenum, and zinc. A polymer or other dust control agent canbe applied to retain the treatment on the seed surface.

Other conventional seed treatment additives include, but are not limitedto, coating agents, wetting agents, buffering agents, andpolysaccharides. At least one agriculturally acceptable carrier can beadded to the seed treatment formulation such as water, solids or drypowders. The dry powders can be derived from a variety of materials suchas wood barks, calcium carbonate, gypsum, vermiculite, talc, humus,activated charcoal, and various phosphorous compounds.

In one embodiment, the seed coating can comprise of at least one filler,which is an organic or inorganic, natural or synthetic component withwhich the active components are combined to facilitate its applicationonto the seed. Preferably, the filler is an inert solid such as clays,natural or synthetic silicates, silica, resins, waxes, solid fertilizers(for example ammonium salts), natural soil minerals, such as kaolins,clays, talc, lime, quartz, attapulgite, montmorillonite, bentonite, ordiatomaceous earths, or synthetic minerals, such as silica, alumina, orsilicates, in particular aluminum or magnesium silicates.

The spore-forming bacterium, optional insect control agent and optionalfungicide control agent, can be combined with any genetically modifiedplant seed capable of germinating to form a plant or plant part that issusceptible to attack by nematodes and/or pathogenic fungi or bacteria.The genetically modified seed can be any seed that results in agenetically modified plant or plant part that expresses insect toxins orherbicide resistance. Further, the genetically modified seed can be anyseed that results in a genetically modified plant or plant part thatexpresses toxins or resistance to bacterial and fungi. Moreover, thegenetically modified seed may be any seed that results in a geneticallymodified plant or plant part that expresses tolerance to environmentalfactors such as water stress and nitrogen production. Suitablegenetically modified seeds include those of cole crops, vegetables,fruits, trees, fiber crops, oil crops, tuber crops, coffee, flowers,legume, cereals, as well as other plants of the monocotyledonous anddicotyledonous species. Preferably, the genetically modified seedsinclude peanut, tobacco, grasses, wheat, barley, rye, sorghum, rice,rapeseed, sugarbeet, sunflower, tomato, pepper, bean, lettuce, potato,and carrot. Most preferably, the genetically modified seeds includecotton, soybean, and corn (sweet, field, seed, or popcorn). Particularlypreferred genetically modified seeds include DELTA AND PINE LAND®glyphosate tolerant and insect tolerant cotton seeds from Monsanto;STONEVILLE™ glyphosate tolerant and insect tolerant cotton seeds fromBayer CropScience; FIBERMAX® glyphosate tolerant and insect tolerantcotton seeds from Bayer CropScience; glyphosate tolerant soybeans fromStine Seed Company; ASGROW® glyphosate tolerant soybean seeds fromMonsanto; PIONEER® glyphosate tolerant and insect tolerant corn seedsfrom DuPont; NORTHRUP KING™ glyphosate tolerant soybean seeds fromSyngenta; glyphosate tolerant and insect tolerant corn seeds from BurrusCompany; and Garst Company (AGRIEDGE™) glyphosate tolerant and insecttolerant corn seeds from Syngenta.

Advantages of the novel combination of spore-forming bacterium andgenetically modified seed will be apparent from the non-limitingexamples below. The following examples demonstrate unexpectedimprovements in overall plant crop yield by combining the spore-formingbacterium with an insect control agent and applying the combination togenetically modified seeds.

Example 1

Example 1 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seeds in this comparison experimentwere sourced from DELTA AND PINE LAND® available from Monsanto, whichare cotton seeds that contains both glyphosate tolerance (Round-upReady® trait) and insect tolerance (Bt gene) gene expressions. Theinsecticide was imidacloprid (Gaucho Grande) or imidacloprid &thiodicarb (AERIS®). The spore-forming bacterium was B. firmus. Thenematode type varied from none, root knot nematode, reiniform nematode,and lance nematode. The concentration of insect control agents was 600gm ai/liter, and in liquid form. The application rate of the insectcontrol agents was 500-1000 gm ai/100 kg. The concentration of B. firmusranged from 100,000 to Ser. No. 10/000,000 colony forming units perseed. The insect control agent and B. firmus were mixed together in anaqueous suspension in enough volume to adequately cover the cottonseed.The rate of the mixture varied from 1369-2608 ml per 100 kg of seed toassure adequate coverage of the various seed sizes. Once planted, thecotton seed grew to full maturity and the results measured in pounds ofcotton per acre. The below table compares the pounds of cotton per acrebetween the control and the control plus spore forming bacterium atvarious nematode types. The yield difference numbers represent anaverage of several experimental results, unless otherwise indicated.

Control + Yield Exp. No. Nematode Control B. firmus difference 1 Lance2957 3205 249 2 Reniform 2044 2132 88 3 Root Knot 2183 2111 −72 4 RootKnot 3226 3298 72 5 Root Knot 3064 3254 190 6 Root Knot 337  300 −37 7Root Knot 1530 1643 113 8 Reniform 1165 1165 0 9 None 2280 2250 −30 10None 2108 2230 122 11 Reniform 2645 2721 76 12 Root Knot 1884 1799 −8513 None 3007 3246 239 14 Root Knot 1615 1602 −13

Some of the negative yield values are the result of uncontrollableenvironmental and planting issues. Regarding Experiment Number 3, thenegative value may be attributable to low planting numbers from plantererror, which will effect overall crop yield. For Experiment No. 6, noreplicates were done because of damage to the planting plots.

Example 2

Example 2 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seed in this comparison experimentwere sourced from STONEVILLE™ available from Bayer CropScience, whichare cotton seeds that contain both glyphosate tolerance (ROUND-UP READY®trait) and insect tolerance (Bt gene) gene expressions. The insecticidewas imidacloprid (GAUCHO GRANDE®) or imidacloprid & thiodicarb (AERIS®).The spore-forming bacterium was B. firmus. The nematode type varied fromnone, root knot nematode and reniform nematode. The concentration ofinsect control agents was 600 gm ai/liter, and in liquid form. Theapplication rate of the insect control agents was 500-1000 gm ai/100 kg.The concentration of B. firmus ranged from 100,000 to Ser. No.10/000,000 colony forming units per seed. The insect control agent andB. firmus were mixed together in an aqueous suspension in enough volumeto adequately cover the cottonseed. The rate of the mixture varied from1369-2608 ml per 100 kg of seed to assure adequate coverage of thevarious seed sizes. Once planted, the cotton seed grew to full maturityand the results measured in pounds of cotton per acre. The below tablecompares the pounds of cotton per acre between the control and thecontrol plus spore forming bacterium at various nematode types. Theyield difference numbers represent an average of several experimentalresults, unless otherwise indicated.

Control + Yield Exp. No. Nematode Control B. firmus difference 1 RootKnot 1658 1728 70 2 Root Knot 2780 2766 −14 3 Reniforms 404 377 −27 4None 596 632 36

Example 3

Example 3 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore-forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seed in this comparison experimentwere sourced from FIBERMAX® available from Bayer CropScience, which arecotton seeds that contains both glyphosate tolerance (ROUND-UP READY®trait) and insect tolerance (Bt gene) gene expressions. The insecticidewas imidacloprid (GAUCHO GRANDE®) or imidacloprid & thiodicarb (AERIS®).The spore-forming bacterium was B. firmus. The nematode types were rootknot nematode and reniform nematode. The concentration of insect controlagents was 600 gm ai/liter, and in liquid form. The concentration of B.firmus ranged from 100,000 to Ser. No. 10/000,000 colony forming unitsper seed. The insect control agent and B. firmus were mixed together inan aqueous suspension in enough volume to adequately cover the cottonseed. The rate of the mixture varied from 1369-2608 ml per 100 kg ofseed to assure adequate coverage of the various seed sizes. Onceplanted, the cotton seed grew to full maturity and the results measuredin pounds of cotton per acre. The below table compares the pounds ofcotton per acre between the control and the control plus spore-formingbacterium at various nematode types. The yield difference numbersrepresent an average of several experimental results, unless otherwiseindicated.

Control + Yield Exp. No. Nematode Control B. firmus difference 1 RootKnot 1541 1589  48 2 Reniform  313  327  14 3 Root Knot 1105 1236 131

Example 4

Example 4 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seed in this comparison experimentare from Stine Seed Company, which are soybean seeds that containglyphosate tolerance (ROUND-UP READY® trait) gene expression. Theinsecticide was imidacloprid (GAUCHO®). The spore-forming bacterium wasB. firmus. The nematode types were none, root knot nematode, and soybeancyst. The concentration of insect control agents was 600 gm ai/liter,and was in liquid form. The concentration of B. firmus ranged from100,000 to Ser. No. 10/000,000 colony forming units per seed. The insectcontrol agent and B. firmus were mixed together in an aqueous suspensionin enough volume to adequately cover the soybean seed. The rate of themixture varied from 261-652 ml per 100 kg of seed to assure adequatecoverage of the various seed sizes. Once planted, the soybean seed grewto full maturity and the results measured in bushels of soybean peracre. The below table compares the bushels of soybean per acre betweenthe control and the control plus spore forming bacterium at variousnematode types. The yield difference numbers represent an average ofseveral experimental results, unless otherwise indicated.

Control + Yield Exp. No. Nematode Control B. firmus difference 1 None64.4 63.4 −1 2 Soybean Cyst 42.8 44 1.2 3 Soybean Cyst 36.1 39.5 3.4 4Soybean Cyst 41 45 4 5 Soybean Cyst 68 67 −1 6 None $9.7 55.3 −4.4 7Root Knot 29.9 31.7 1.8 8 None 53 53.3 0.3

Example 5

Example 5 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore-forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seed in this comparison experimentwas ASGROW® from Monsanto, which is a soybean seed that containsglyphosate tolerance (ROUND-UP READY® trait) gene expression. Theinsecticide was imidacloprid (GAUCHO®). The spore-forming bacterium wasB. firmus. The nematode types were root knot nematode and soybean cyst.The concentration of insect control agent was 600 gm ai/liter and werein liquid form. The concentration of B. firmus ranged from 100,000 toSer. No. 10/000,000 colony forming units per seed. The insect controlagent and B. firmus were mixed together in an aqueous suspension inenough volume to adequately cover the soybean seed. The rate of themixture varied from 261-652 ml per 100 kg of seed to assure adequatecoverage of the various seed sizes. Once planted, the soybean seed grewto full maturity and the results measured in bushels of soybean peracre. The below table compares the bushels of soybean per acre betweenthe control and the control plus spore forming bacterium. The yielddifference numbers represent an average of several experimental results,unless otherwise indicated.

Control + Yield Exp. No. Nematode Control B. firmus difference 1 RootKnot and 25.4 26.9 1.5 Soybean Cyst

Example 6

Example 6 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore-forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seed in this comparison experimentwere sourced from PIONEER® from DuPont, which are soybean seeds thatcontain glyphosate tolerance (ROUND-UP READY® trait) gene expression.The insecticide was imidacloprid (GAUCHO®). The spore-forming bacteriumwas B. firmus. The nematode types were none and soybean cyst. Theconcentration of insect control agent was 600 gm ai/liter, and were inliquid form. The concentration of B. firmus ranged from 100,000 to Ser.No. 10/000,000 colony forming units per seed. The insect control agentand B. firmus were mixed together in an aqueous suspension in enoughvolume to adequately cover the soybean seed. The rate of the mixturevaried from 261-652 ml per 100 kg of seed to assure adequate coverage ofthe various seed sizes. Once planted, the soybean seed grew to fullmaturity and the results measured in bushels of soybean per acre. Thebelow table compares the bushels of soybean per acre between the controland the control plus spore-forming bacterium. The yield differencenumbers represent an average of several experimental results, unlessotherwise indicated.

Control + Yield Exp. No. Nematode Control B. firmus difference 1 None61   62.6 1.6 2 Soybean Cyst 27.2 28.3 1.1 3 Soybean Cyst 29.9 31.7 1.8

Example 7

Example 7 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore-forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seed in this comparison experimentwas sourced from NORTHRUP KING™ of Syngenta Seeds, which is a soybeanseed that contains glyphosate tolerance (ROUND-UP READY® trait) geneexpression. The insecticide was imidaloprid (GAUCHO®). The spore-formingbacterium was B. firmus. The nematode type was soybean cyst. Theconcentration of insect control agent was 600 gm ai/liter, and was inliquid form. The concentration of B. firmus ranged from 100,000 to Ser.No. 10/000,000 colony forming units per seed. The insect control agentand B. firmus were mixed together in an aqueous suspension in enoughvolume to adequately cover the soybean seed. The rate of mixture seedcoverage varied from 261-652 ml per 100 kg of seed. Once planted, thesoybean seed grew to full maturity and the results measured in bushelsof soybean per acre. The below table compares the bushels of soybean peracre between the control and the control plus spore forming bacterium.The yield difference numbers represent an average of severalexperimental results, unless otherwise indicated.

Control + Yield Exp. No. Nematode Control B. firmus difference 1 SoybeanCyst 44 45.3 1.3 2 Soybean Cyst 46 46.3 0.3

Example 8

Example 8 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore-forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seed in this comparison experimentwas sourced from PIONEER® from DuPont, which is a corn seed thatcontains both glyphosate tolerance (ROUND-UP READY® trait) and insecttolerance (Bt gene) gene expressions. The insecticide was clothianidin(PONCHO®). The spore-forming bacterium was B. firmus. Multiple speciesof nematodes were present. The concentration of insect control agent was600 gm ai/liter, and was in liquid form. The concentration of B. firmusranged from 100,000 to Ser. No. 10/000,000 colony forming units perseed. The insect control agent and B. firmus were mixed together in anaqueous suspension in enough volume to adequately cover the corn seed.The rate of the mixture varied from 522-1044 ml per 100 kg of seed toassure adequate coverage of the various seed sizes. Once planted, thecorn seed grew to full maturity and the results measured in bushels ofcorn per acre. The below table compares the bushels of corn per acrebetween the control and the control plus spore forming bacterium. Theyield difference numbers represent an average of several experimentalresults, unless otherwise indicated.

Control + Yield Exp. No. Nematode Control B. Firmas difference 1Multiple species 119.3 132.3 13

Example 9

Example 9 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore-forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seeds in this comparison experimentwere sourced from Burrus Company, which are corn seeds that containsboth glyphosate tolerance (ROUND-UP READY® trait) and insect tolerance(varies) gene expressions. The insecticide was clothiainidin (PONCHO®).The spore-forming bacterium was B. firmus. Multiple species of nematodeswere present. The concentration of insect control agent was 600 gmai/liter, and in liquid form. The concentration of B. firmus ranged from100,000 to Ser. No. 10/000,000 colony forming units per seed. The insectcontrol agent and B. firmus were mixed together in an aqueous suspensionin enough volume to adequately cover the corn seed. The rate of themixture varied from 522-1044 ml per 100 kg of seed to assure adequatecoverage of the various seed sizes. Once planted, the corn seed grew tofull maturity and the results measured in bushels of corn per acre. Thebelow table compares the bushels of corn per acre between the controland the control plus spore-forming bacterium. The yield differencenumbers represent an average of several experimental results, unlessotherwise indicated.

Insect Control + Yield Exp. No. Nematode tolerance gene Control B.firmus difference 1 Multiple Corn root 148.1 146.8 −1.3 species worm andBt 2 Multiple Bt 173   183.5 11.5 species

Example 10

Example 10 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore-forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seed in this comparison experimentwas sourced from Garst Company (AGRIEDGE™) of Syngenta Seeds, which is acorn seed that contains both glyphosate tolerance (ROUND-UP READY®trait) and insect tolerance (corn root worm) gene expressions. Theinsecticide was clothianidin (PONCHO®). The spore-forming bacterium wasB. firmus. Multiple species of nematodes were present. The concentrationof insect control agent was 600 gm ai/liter, and in liquid form. Theconcentration of B. firmus ranged from 100,000 to Ser. No. 10/000,000colony forming units per seed. The insect control agent and B. firmuswere mixed together in an aqueous suspension in enough volume toadequately cover the corn seed. The rate of the mixture varied from522-1044 ml per 100 kg of seed to assure adequate coverage of thevarious seed sizes. Once planted, the corn seed grew to full maturityand the results measured in bushels of corn per acre. The below tablecompares the bushels of corn per acre between the control and thecontrol plus spore-forming bacterium. The yield difference numbersrepresent an average of several experimental results, unless otherwiseindicated.

Control + Yield Exp. No. Nematode Control B. firmus difference 1Multiple species 170.12 363.3 −6.82 2 Multiple species 136 133.1 −2.9

The negative yield difference values for each experiment are most likelyattributable to damages planting plots, which resulted in no replicateexperiments. When averaging Experiment Number 1 and 2, the Control is142.6, the Control+B. firmus is 146.1, and the Yield difference is 3.5.Therefore, the use of the spore-forming bacterium shows a net positiveresult.

Example 11

Example 11 shows the crop yield results under nematode pressure for thecombination of spore-forming bacterium and insect control agent appliedto genetically modified seed (control+spore-forming bacterium) comparedto genetically modified seed with just an insect control agent(control). The genetically modified seed in this comparison experimentwas an unknown corn seed purchased from a production company thatcontains various gene expressions for both herbicide resistance andinsect resistance. The insecticide was clothianidin (PONCHO®). Thespore-forming bacterium was B. firmus. Multiple species of nematodeswere present. The concentration of insect control agent was 600 gmai/liter, and in liquid form. The concentration of B. firmus ranged from100,000 to Ser. No. 10/000,000 colony forming units per seed. The insectcontrol agent and B. firmus were mixed together in an aqueous suspensionin enough volume to adequately cover the corn seed. The rate of themixture varied from 522-1044 ml per 100 kg of seed to assure adequatecoverage of the various seed sizes. Once planted, the corn seed grew tofull maturity and the results measured in bushels of corn per acre. Thebelow table compares the bushels of corn per acre between the controland the control plus spore-forming bacterium. The yield differencenumbers represent an average of several experimental results, unlessotherwise indicated.

Gene Control + Yield Exp. No. Nematode Expression Control B. firmusdifference 1 Multiple Bt and corn 131.7 146   14.3 species borer 2Multiple Stand 194.8 200.5  5.7 species Hurculex 3 Multiple Bt and 204.2227.6 23.4 species Hurculex

The results above are surprising. As shown above, most seed varietiesfaired better with the combination of spore-forming bacterium and insectcontrol agent verses just the insect control agent.

According to some embodiments, the following percent crop yieldincreases were obtained. Regarding soybean seed, the addition ofspore-forming bacterium to NORTHUP KING™ soybean seed, PIONEER® soybeanseed, and soybean seed from Stine Seed Company showed about a 2%increase, 5%-10% increase, and 4.3% increase, respectively, in soybeanbushel yield. This improvement, spread out over several hundred acresresults in a significant improvement in crop yield. Regarding cottonseed, the addition of spore-forming bacterium to DELTA AND PINE LAND®cotton seed and FIBERMAX® cotton seed showed about a 3% increase and6.5% increase, respectively, in pounds of cotton per acre. Again,spreading this improvement over several hundred acres of cotton plantyields a drastic improvement in yield. Regarding corn seed, the additionof spore-forming bacterium to corn seed from Burrus Seed Company andunknown commercially available corn seed showed about a 3.2% increaseand 8% increase, respectively, in bushels of corn per acre. Again, thisresults in a huge improvement in overall crop yield when spread out overseveral hundred acres. Overall, there is a surprising advantage in cropyield when using the combination of spore-forming bacterium and insectcontrol agent with genetically modified seeds.

Additional advantages, features and modifications will readily occur tothose skilled in the art. Therefore, the invention in its broaderaspects is not limited to the specific details, and representativedevices, shown and described herein. Accordingly, various modificationsmay be made without departing from the spirit or scope of the generalinventive concept as defined by the appended claims and theirequivalents.

All documents referred to herein are specifically incorporated herein byreference in their entireties.

As used herein and in the following claims, articles such as “the”, “a”and “an” can connote the singular or plural.

1. A product comprising: at least one spore-forming bacterium;optionally, at least one insect control agent; and at least onegenetically modified seed, plant, or plant part.
 2. The product of claim1, wherein said product further comprises at least one insect controlagent and at least one genetically modified seed or plant.
 3. Theproduct of claim 2, wherein the insect control agent is at least onesystemic neonicotinoid insecticide.
 4. The product of claim 3, whereinthe at least one neonicotinoid insecticide is selected from the groupconsisting of1-(6-chloro-3-pyridylmethyl)-N-nitroimidazolidin-2-ylideneamine(imidacloprid),3-(6-chloro-3-pyridylmethyl)-1,3-thiazolidin-2-ylidenecyanamide(thiacloprid),1-(2-chloro-1,3-thiazol-5-ylmethyl)-3-methyl-2-nitroguanidine(clothianidin), nitempyran,N.sup.1-[(6-chloro-3-pyridyl)methyl]-N.sup.2-cyano-N.sup.1-methylacetamidine(acetamiprid),3-(2-chloro-1,3-thiazol-5-ylmethyl)-5-methyl-1,3,5-oxadiazinan-4-ylidene(nitro)amine(thiamethoxam) and1-methyl-2-nitro-3-(tetrahydro-3-furylmethyl)guanidine (dinotefuran). 5.The product of claim 2, wherein the at least one spore-forming bacteriumexhibits nematicidal activity.
 6. The product of claim 2, wherein saidgenetically modified seed is insect tolerant.
 7. The product of claim 2,wherein said genetically modified seed is transformed with a gene fromB. thuringiensis.
 8. The product of claim 2, wherein said geneticallymodified seed is glyphosate tolerant.
 9. The product of claim 2, whereinsaid genetically modified seed is both insect tolerant and glyphosatetolerant.
 10. The product of claim 2, wherein said genetically modifiedseed is selected from the group consisting of: DELTA AND PINE LAND®glyphosate tolerant and insect tolerant cotton seeds; STONEVILLE™glyphosate tolerant and insect tolerant cotton seeds; FIBERMAX®glyphosate tolerant and insect tolerant cotton seeds; ASGROW® glyphosatetolerant soybean seeds; PIONEER® glyphosate tolerant and insect tolerantcorn seeds; NORTHRUP KING™ glyphosate tolerant soybean seeds; andAGRIEDGE™ glyphosate tolerant and insect tolerant corn seeds fromSyngenta.
 11. The product of claim 5, wherein the at least onespore-forming bacterium is selected from the group consisting ofBacillus amyloliquefaciens, Bacillus firmus, Bacillus subtillis, andBacillus pumulis.
 12. The product of claim 11, wherein the Bacillusfirmus comprises strain CNCM I-1582.
 13. The product of claim 11,wherein the Bacillus amyloliquefaciens comprises strain IN37a.
 14. Theproduct of claim 11, wherein the Bacillus subtilis comprises strainGB03.
 15. The product of claim 11, wherein the Bacillus pumuliscomprises strain GB34.
 16. The product of claim 1, further comprising atleast one additional chemical fungicide.
 17. A method of protecting agenetically modified seed, plant, or plant part from nematodes,comprising applying to the seed, plant, or plant part at least onespore-forming bacterium; and, optionally, at least one insect controlagent.
 18. The method of claim 17, wherein said spore-forming bacteriumand optional insect control agent are applied simultaneously to saidgenetically modified seed, plant, or plant part.
 19. The method of claim17, wherein said genetically modified seed, plant, or plant part isinsect tolerant.
 20. The method of claim 17, wherein said geneticallymodified seed, plant, or plant part is glyphosate tolerant.
 21. Themethod of claim 17, wherein said genetically modified seed, plant, orplant part is transformed with a gene from B. thuringiensis.
 22. Themethod of claim 17, wherein said at least one spore-forming bacterium isselected from the group consisting of Bacillus amyloliquefaciens,Bacillus firmus, Bacillus subtillis, and Bacillus pumulis.
 23. A methodof protecting a genetically modified seed, plant, or plant part fromnematodes comprising providing at least one composition comprising0.0001 to 20% by weight of at least one spore-forming bacterium and0.001 to 20% by weight of at least one insect control agent; andapplying the composition to the seed, plant, or plant part.
 24. Themethod of claim 23, wherein the composition is applied by a methodselected from the group consisting of: drip irrigation, sprinklers,foliar spray, seed coating, soil injection or soil drenching.
 25. Acomposition for protecting a genetically modified seed, plant, or plantpart from nematodes comprising: (i) at least one spore-forming bacteriumin an amount of from about 2% by weight to 80% by weight; (ii) at leastone insect control agent in an amount of from about 1% by weight toabout 80% by weight; and (iii) a solvent.
 26. The composition of claim29, wherein said spore-forming bacterium is selected from the groupconsisting of: Bacillus amyloliquefaciens, Bacillus firmus, Bacillussubtillis, and Bacillus pumulis.
 27. A method of manufacturing agenetically modified seed treated with at least one spore-formingbacteria and an optional insect control agent comprising: (i) applyingsaid spore-forming bacteria and optional insect control agent to saidgenetically modified seed; and (ii) mixing said genetically modifiedseed to achieve a substantially uniform treatment.
 28. The method ofclaim 27, wherein a continuous coating machine is used to apply saidspore-forming bacteria and optional insect control agent.
 29. The methodof claim 28, wherein a computer system monitors the flow of geneticallymodified seed into the continuous coating machine.
 30. The method ofclaim 27, wherein a batch coating process is used to apply saidspore-forming bacteria and optional insect control agent.
 31. The methodof claim 30, wherein said batch coating process comprises: (i) weighinga set amount of genetically modified seed; (ii) placing the seed into aclosed treating chamber; (iii) adding the spore-forming bacteria andoptional insect control agent into the treating chamber; and (iv) mixingsaid genetically modified seed, spore-forming bacteria, and optionalinsect control agent.